Multipotent adult stem cells having an ability of oct4 expression derived from umbilical cord blood and method for preparing the same

a multi-potent stem cell, umbilical cord blood technology, applied in the field of multi-potent stem cells, can solve the problems of difficult to define these cells as true multi-potent stem cells, many problems in the file of organ transplantation, etc., and achieve the effect of positive or negative immunological responses

Inactive Publication Date: 2009-12-10
SEOUL NAT UNIV R&DB FOUND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0025]In the present invention, the adult stem cells preferably additionally show positive immunological responses to one or more selected from the group consisting of SH-2, SH-3 and SH-4, and preferably show positive or negative immunological responses to CD44, CD105 and CD117. Moreover, the cells derived from mesoderm are preferably osteogenic cells or endothelial cells, and the cells derived from ectoderm are preferably nerve cells.
[0026]Another features and embodiments of the present invention will be more clarified from the following “detailed description” and the appended “claims”.

Problems solved by technology

Moreover, there are many problems in the filed of organ transplantation.
These cells are called “embryonic stem cells” and can differentiate into various other tissue cells but cannot form new living organisms.
However, because the stem cells are capable of differentiating into a limited variety of cell types, including differentiation into osteocytes or skeletal muscles by mesenchymal stem cells, differentiation into heart cells by heart stem cells, and differentiation into vascular endothelial cells by endothelial progenitor cells, it is difficult to define these cells as true multipotent stem cells.

Method used

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  • Multipotent adult stem cells having an ability of oct4 expression derived from umbilical cord blood and method for preparing the same
  • Multipotent adult stem cells having an ability of oct4 expression derived from umbilical cord blood and method for preparing the same
  • Multipotent adult stem cells having an ability of oct4 expression derived from umbilical cord blood and method for preparing the same

Examples

Experimental program
Comparison scheme
Effect test

example 1

Isolation of Adult Stem Cells from Umbilical Cord Blood

[0045]Umbilical cord blood was collected from full-term and preterm newborns in Seoul National University Hospital and Samsung Cheil Hospital according to Institutional Review Board guidelines.

[0046]70-100 ml of the collected umbilical cord blood sample was diluted in PBS at a ratio of 1:1 to stir. Then, the blood sample was separated on ficoll at a ratio of 15:25, in which the blood sample (diluted in PBS at a ratio of 1:1) was spilled smoothly onto 15 ml of Ficoll solution to cause layer separation, followed by centrifugation at 2500 rpm for 20 minutes. After the centrifugation, three different layers were formed starting from the bottom, among them, buffy coat (the middle layer:monocytic layer) was taken with a micropipette, and washed three times with HBSS, followed by centrifugation at 1800 rpm for 15 minutes, from which upper layer is completely removed and a precipitation was stored in ice.

[0047]The precipitation was shak...

example 2

Immunological Characteristics of Umbilical Cord Blood-Derived Multipotent Adult Stem Cells

[0050]To examine the immunological characteristics of the umbilical cord blood-derived multipotent adult stem cells obtained in Example 1, the pattern of cell surface antigen expression was analyzed. For this purpose, 2×106-107 of the cells cultured in Example 1 were washed with PBS solution and incubated with their corresponding antibodies at room temperature. The expression or non-expression of the antigens was analyzed with a flow cytometer. Also, PAS staining periodic acid Schiff staining) was conducted.

[0051]As a result, as shown in FIG. 4, the inventive umbilical cord blood-derived multipotent adult stem cells showed 63.38%, 96.54% and 63.99% of positive responses to CD45, SH-2 and SH-3, respectively, and more than 90% of negative response to CD34. Also, immunophenotypes of other antigens were analyzed and as a result, the immunological characteristics of the multipotent adult stem cells ...

example 3

Differentiation of Umbilical Cord Blood-Derived Multipotent Adult Stem Cells into Osteogenic Cells

[0053]The umbilical cord blood-derived multipotent adult stem cell broth obtained in Example 1 was diluted in 1 ml of osteogenesis-inducing medium (0.1 μmol / L dexamethasone (Sigma, USA), 0.05 mmol / L ascorbic acid-2-phosphate (Sigma, USA), 10 mmol / L beta-glycophosphate (Sigma), and 5-30% human serum or plasma) to count the number of cells. Then, the cells were cultured in a flask (5% CO2; 37° C.; medium replaced one time at 3-4-day intervals) to induce the differentiation of the multipotent adult stem cells into osteogenic cells. 14 days after the initiation of the culture, it was confirmed by Von-Kassa staining that the umbilical cord blood-derived multipotent adult stem cells differentiated into osteogenic cells (see FIG. 4).

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Abstract

The present invention relates to multipotent adult stem cells expressing Oct4, derived from umbilical cord blood (UCB) and also these cell are expressing CD29, CD31, CD44, simultaneously, a method for preparing the same, and more specifically to multipotent adult stem cells which are obtained by culturing umbilical cord blood-derived monocytes in a medium containing bFGF (basic fibroblast growth factor) and human serum or plasma. In addition, multipotent adult stem cells expressing Oct-4 from UCB are morphologically spindle or round shaped cells Although the stem cells according to the present invention are adult stem cells, they are multipotent and capable of differentiating into ectodermal-, messodermal-, endodermal-originated tissue or cells including osteogenic cells or nerve cells etc, thus they can be effectively used in the treatment of intractable diseases and incurable diseases.

Description

TECHNICAL FIELD[0001]The present invention relates to multipotent adult stem cells expressing Oct4 from umbilical cord blood and a method for preparing the same, and more specifically, to multipotent adult stem cells which are obtained by culturing umbilical cord blood-derived monocyte in a medium containing bFGF (basic fibroblast growth factor) and human serum or plasma to isolate.BACKGROUND ART[0002]Although many diseases of the past have been cured in our modern society due to the development of life sciences and medical sciences, there still have been inveterate diseases or incurable diseases such as ischemic necrosis, cancer, dementia and a severe burn, and the like. Moreover, there are many problems in the filed of organ transplantation. As a therapy to treat the root cause of these diseases, cell therapy has been in the limelight these days.[0003]Cell therapy is a method for treating or preventing diseases by externally proliferating or sorting autologous stem cells, allogeni...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12N5/06C12N5/074
CPCC12N2501/115C12N5/0607C12N5/0647
Inventor KANG, KYUNG SUN
Owner SEOUL NAT UNIV R&DB FOUND
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